Purification to homogeneity of GD3 synthase and partial purification of GM3 synthase from rat brain.
A CMP-sialic acid: GM3 sialyltransferase (GD3 synthase) and a CMP-sialic acid: LacCer sialyltransferase (GM3 synthase) have been purified 10,000- and 3,000-fold, respectively, from the Triton X-100 extract of rat brain. The two enzymes were purified and resolved by affinity chromatography on two successive CDP-Sepharose columns by NaCl gradient elution. Final purification of GD3 synthase was achieved by specific elution from a 'GM3 acid'-Sepharose column with buffer containing GM3. Sodium dodecylsulfate-gel electrophoresis of GD3 synthase revealed a single major protein band with an apparent molecular weight of 55,000.
Gu
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XB
Department of Biochemistry and Molecular Biophysics, Virginia Commonwealth University, Medical College of Virginia, Richmond 23298-0614.
Gu
T J
TJ
Yu
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RK
D000818:Animals; D001921:Brain / Q000201:enzymology*; D002846:Chromatography, Affinity / Q000379:methods; D003902:Detergents / Q000494:pharmacology; D004789:Enzyme Activation; D007700:Kinetics; D008861:Microsomes / Q000201:enzymology*; D008970:Molecular Weight; D009928:Organ Specificity; D051381:Rats; D012799:Sialyltransferases / Q000302:isolation & purification* / Q000378:metabolism
D003902:Detergents; D012799:Sialyltransferases; C043662:alpha-N-acetylneuraminate alpha-2,8-sialyltransferase; C021343:haematoside synthetase